Göttgens B, Barton L M, Gilbert J G, Bench A J, Sanchez M J, Bahn S, Mistry S, Grafham D, McMurray A, Vaudin M, Amaya E, Bentley D R, Green A R, Sinclair A M
University of Cambridge, Department of Haematology, MRC Centre, Hills Road, Cambridge CB2 2QH, UK.
Nat Biotechnol. 2000 Feb;18(2):181-6. doi: 10.1038/72635.
The SCL gene encodes a highly conserved bHLH transcription factor with a pivotal role in hemopoiesis and vasculogenesis. We have sequenced and analyzed 320 kb of genomic DNA composing the SCL loci from human, mouse, and chicken. Long-range sequence comparisons demonstrated multiple peaks of human/mouse homology, a subset of which corresponded precisely with known SCL enhancers. Comparisons between mammalian and chicken sequences identified some, but not all, SCL enhancers. Moreover, one peak of human/mouse homology (+23 region), which did not correspond to a known enhancer, showed significant homology to an analogous region of the chicken SCL locus. A transgenic Xenopus reporter assay was established and demonstrated that the +23 region contained a new neural enhancer. This combination of long-range comparative sequence analysis with a high-throughput transgenic bioassay provides a powerful strategy for identifying and characterizing developmentally important enhancers.
SCL基因编码一种高度保守的bHLH转录因子，在造血和血管生成中起关键作用。我们已对构成人类、小鼠和鸡的SCL基因座的320 kb基因组DNA进行了测序和分析。长程序列比较显示了人类/小鼠同源性的多个峰值，其中一部分与已知的SCL增强子精确对应。哺乳动物和鸡序列之间的比较确定了一些但不是所有的SCL增强子。此外，一个人类/小鼠同源性峰值（+23区域），它与已知的增强子不对应，与鸡SCL基因座的一个类似区域显示出显著的同源性。建立了一种转基因非洲爪蟾报告基因检测方法，并证明+23区域包含一个新的神经增强子。这种长程比较序列分析与高通量转基因生物检测相结合的方法，为鉴定和表征发育上重要的增强子提供了一种强大的策略。
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